CN114426938A

CN114426938A - Lactobacillus plantarum CCFM1202 with PCOS (prestressed concrete cylinder pipes) relieving function and application thereof

Info

Publication number: CN114426938A
Application number: CN202210120743.7A
Authority: CN
Inventors: 王刚; 贺禹丰; 赵建新; 张灏; 陈卫
Original assignee: Jiangnan University
Current assignee: Jiangnan University
Priority date: 2022-02-08
Filing date: 2022-02-08
Publication date: 2022-05-03
Anticipated expiration: 2042-02-08
Also published as: CN114426938B

Abstract

The invention discloses lactobacillus plantarum CCFM1202 with a PCOS (prestressed concrete) relieving function and application thereof, belonging to the technical field of functional microorganisms. The lactobacillus plantarum CCFM1202 provided by the invention has good tolerance to gastrointestinal fluids, and can be used for remarkably improving abnormal testosterone level, abnormal luteinizing hormone level, abnormal triglyceride level and insulin resistance caused by polycystic ovary syndrome; remarkably recovering the reduction of the diversity of intestinal flora caused by polycystic ovarian syndrome; in addition, the lactobacillus plantarum CCFM1202 can relieve the reproductive cycle disorder caused by polycystic ovarian syndrome, the estrus ratio is reduced to 23.33 +/-8.17%, and the difference with a normal group is not obvious. Meanwhile, the bisphenol F adsorbent has strong bisphenol F in-vitro adsorption capacity.

Description

Lactobacillus plantarum CCFM1202 with PCOS relieving function and application thereof

Technical Field

The invention relates to lactobacillus plantarum CCFM1202 with a function of relieving PCOS and application thereof, belonging to the technical field of functional microorganisms.

Background

In recent years, economic development causes changes of life styles and unreasonable dietary structures of people in China, and the prevalence rate of polycystic ovary syndrome (PCOS) of women of childbearing age is greatly increased. Statistics by the National Institutes of Health (NIH) show that 4-10% of women of reproductive age worldwide suffer from polycystic ovarian syndrome. Ovulation failure due to polycystic ovarian syndrome seriously threatens the fertility of women, and the cost for diagnosing and treating the disease is over $ 1000 billion per year around the world. Therefore, the global burden of addressing polycystic ovarian syndrome has been at hand.

The etiology of polycystic ovarian syndrome is not clear, and the pathogenesis of polycystic ovarian syndrome is related to abnormal sex hormone level, abnormal metabolism (such as insulin resistance) and local ovarian lesion. Typical symptoms of polycystic ovarian syndrome include abnormal menstruation, hyperandrogenism, and ovarian polycystic lesions. Furthermore, polycystic ovarian syndrome is accompanied by a disturbance of the intestinal microecology. Meanwhile, the patient also shows emotional adverse reaction due to the high androgen and hirsutism caused by the high androgen. Bisphenol F is a common environmental pollutant and has close relation with abnormal sex hormone level and the onset of polycystic ovary syndrome.

Disclosure of Invention

This section is for the purpose of summarizing some aspects of embodiments of the invention and to briefly introduce some preferred embodiments. In this section, as well as in the abstract and the title of the invention of this application, simplifications or omissions may be made to avoid obscuring the purpose of the section, the abstract and the title, and such simplifications or omissions are not intended to limit the scope of the invention.

In order to overcome the defects in the prior art, the invention provides a Lactobacillus plantarum (Lactobacillus plantarum) CCFM1202 which is preserved in Guangdong province microbial strain preservation center in 12 and 17 months in 2021, the preservation address is Guangzhou city Mr. No. 59 large institute of Michelia Tokyo No. 5 building Guangdong province microbial research institute of Michelia Tokyo No. 100, the preservation number is GDMCC No: 62137.

the invention also provides a probiotic preparation containing the lactobacillus plantarum CCFM 1202.

In one embodiment, the Lactobacillus plantarum CCFM1202 content of the probiotic formulation is ≥ 1 × 10⁶CFU/mL or more than or equal to 1X 10⁶CFU/g。

The invention also provides a pharmaceutical composition containing the lactobacillus plantarum CCFM 1202.

In one embodiment, the pharmaceutical composition further comprises a pharmaceutical carrier and/or a pharmaceutical excipient.

The invention also provides a fermented food which is prepared by fermenting and producing the lactobacillus plantarum CCFM 1202.

In one embodiment, the fermented food product comprises a solid food product, a liquid food product, a semi-solid food product.

In one embodiment, the fermented food product comprises dairy products, bean products, fruit and vegetable products, and the dairy products comprise milk, sour cream and cheese; the fruit and vegetable products comprise cucumber, carrot, beet, celery and cabbage products.

The invention also provides application of the lactobacillus plantarum CCFM1202 in preparation of a product for relieving polycystic ovary syndrome.

In one embodiment, the product has at least one of the following effects:

(1) the compound has better tolerance to simulated gastrointestinal fluid;

(2) restoring the reproductive cycle disorder of the polycystic ovarian syndrome rat;

(3) significantly reducing the testosterone level of a rat with polycystic ovarian syndrome;

(4) significantly reducing the level of luteinizing hormone of a rat with polycystic ovarian syndrome;

(5) significantly reducing the level of triglyceride of a rat with polycystic ovarian syndrome;

(6) significantly reducing the insulin resistance index of the rat with polycystic ovarian syndrome;

(7) obviously improving the reduction of diversity of intestinal flora of the rat with the polycystic ovarian syndrome;

(8) has obvious in-vitro adsorption capacity to bisphenol F.

In one embodiment, the product contains Lactobacillus plantarum CCFM1202 in an amount of 1 × 10 or more⁶CFU/mL or more than or equal to 1X 10⁶CFU/g。

In one embodiment, the product comprises a food or pharmaceutical product.

In one embodiment, the medicament comprises lactobacillus plantarum CCFM1202, a pharmaceutical carrier and/or a pharmaceutical excipient.

In one embodiment, the drug carrier comprises a microcapsule, microsphere, nanoparticle, and/or liposome.

In one embodiment, the pharmaceutical product is in the form of a powder, granule, capsule, tablet, pill, or oral liquid.

In one embodiment, the food product comprises a solid food product, a liquid food product, a semi-solid food product.

In one embodiment, the food product comprises dairy products, soy products, fruit and vegetable products.

The invention has the beneficial effects that:

the lactobacillus plantarum CCFM1202 can obviously improve the abnormal testosterone level, the abnormal luteinizing hormone level, the abnormal triglyceride level and the insulin resistance caused by the polycystic ovary syndrome; remarkably recovering the reduction of the diversity of intestinal flora caused by polycystic ovarian syndrome; the lactobacillus plantarum CCFM1202 can relieve the reproductive cycle disorder caused by polycystic ovarian syndrome, the estrus ratio is reduced to 23.33 +/-8.17%, and the difference with a normal group is not obvious. In addition, the lactobacillus plantarum CCFM1202 has strong adsorption capacity to bisphenol F, has the capacity of relieving the toxicity of the bisphenol F, and the in vitro adsorption rate reaches 60 percent in 24 hours. The lactobacillus plantarum CCFM1202 can well tolerate simulated gastrointestinal fluid, can be used for preparing pharmaceutical compositions and fermented foods for relieving polycystic ovarian syndrome, improving intestinal flora, recovering physiological cycle and adsorbing bisphenol F, and has very wide application prospects.

Proof of biological preservation

Lactobacillus plantarum (Lactobacillus plantarum) CCFM1202, which was deposited at the Guangdong province microbial strain collection center at 12 months and 17 days 2021, the collection address is the Guangdong province microbial research institute of No. 59, No. 5, No. 59, Mieli Zhou, Middu, Guangzhou city, and the collection number is GDMCC No: 62137.

drawings

In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings needed to be used in the description of the embodiments will be briefly introduced below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and it is obvious for those skilled in the art to obtain other drawings based on these drawings without inventive exercise.

Wherein:

FIG. 1 shows the colony morphology of Lactobacillus plantarum CCFM 1202;

FIG. 2 is a graph showing the effect of Lactobacillus plantarum CCFM1202 on serum Testosterone (Testosterone) levels in polycystic ovary syndrome rats;

FIG. 3 is a graph of the effect of Lactobacillus plantarum CCFM1202 on the serum Luteinizing Hormone (LH) level in polycystic ovary syndrome rats;

FIG. 4 is a graph of the effect of Lactobacillus plantarum CCFM1202 on serum Triglyceride (TG) levels in rats with polycystic ovarian syndrome;

FIG. 5 is a graph of the effect of Lactobacillus plantarum CCFM1202 on the level of the insulin resistance index (HOMA-IR) in polycystic ovary syndrome rats;

FIG. 6 is a graph of the effect of Lactobacillus plantarum CCFM1202 on the Shannon index (Shannon) of the intestinal flora of polycystic ovarian syndrome rat;

FIG. 7 shows the in vitro adsorption of bisphenol F by Lactobacillus plantarum CCFM 1202; note: indicates that the experimental group had significant differences (p <0.05) compared to Letrozole model (Letrozole). a and b indicate that the groups represented by different letters have significant difference (p < 0.05).

Detailed Description

In order to make the aforementioned objects, features and advantages of the present invention comprehensible, embodiments accompanied with examples are described in detail below.

In the following description, numerous specific details are set forth in order to provide a thorough understanding of the present invention, but the present invention may be practiced in other ways than those specifically described and will be readily apparent to those of ordinary skill in the art without departing from the spirit of the present invention, and therefore the present invention is not limited to the specific embodiments disclosed below.

Furthermore, reference herein to "one embodiment" or "an embodiment" means that a particular feature, structure, or characteristic described in connection with the embodiment is included in at least one implementation of the invention. The appearances of the phrase "in one embodiment" in various places in the specification are not necessarily all referring to the same embodiment, nor are separate or alternative embodiments mutually exclusive of other embodiments.

Lactobacillus plantarum CCFM1202 has the following biological properties:

(1) the characteristics of the thallus are as follows: gram-positive, non-sporulating, immotile bacteria.

(2) Colony characteristics: aerobic or anaerobic culture for 36 hr to form obvious colony of 0.5-2mm diameter, round front shape, raised side shape, regular edge, creamy white color, opacity, moist and smooth surface, and no pigment generation, as shown in figure 1.

(3) Growth characteristics: under aerobic or anaerobic conditions at a constant temperature of 37 ℃, the medium was cultured in mrss medium for about 16 hours to the end of log.

(4) The compound has better tolerance to simulated gastrointestinal fluid;

(5) can obviously improve the abnormal reproductive cycle of the rat with the polycystic ovarian syndrome;

(6) can obviously improve the abnormal testosterone level of the rat with the polycystic ovarian syndrome;

(7) can obviously improve the abnormal level of luteinizing hormone of a rat with polycystic ovarian syndrome;

(8) can obviously improve the abnormal level of the triglyceride of a rat with the polycystic ovarian syndrome;

(9) can obviously improve the insulin resistance of the rat with the polycystic ovarian syndrome;

(10) can obviously improve the reduction of diversity of intestinal flora of the rat with the polycystic ovarian syndrome;

(11) the in vitro adsorption capacity to bisphenol F is remarkable;

the extraction method of the strain comprises the following steps:

separation and screening of lactic acid bacteria

(l) 1g of fresh faeces from healthy persons were taken. Enriching the sample in a culture medium containing sorbitol GM17 at 35 ℃ for 12 h;

(2) performing gradient dilution on the enriched sample, then coating the enriched sample on a GM17 solid plate added with 0.02% of olcresol purple, and culturing for 24-48 h;

(3) selecting single bacterial colony with obvious color changing circle and according with the basic morphology of lactobacillus, carrying out plate streaking purification, and screening and separating out lactobacillus;

(4) and culturing the single colony in a liquid GM17 culture solution for 24h, performing gram staining, and selecting gram-positive bacteria for subsequent tests.

(II) preliminary identification of lactic acid bacteria for fermentation: caldolytic ring assay

(l) Culturing the lactic acid bacteria obtained by screening in the step (I) in a liquid sorbitol GM17 culture solution for 24h, and then centrifuging l mL of culture at 8000rpm for 2 min;

(2) with 0.05M KH₂PO₄Washing the solution twice;

(3) resuspending the resulting pellet and streaking on sorbitol GM 17-0.75% CaCO₃Culturing for 24 hours on the solid culture medium;

(4) selecting bacterial colonies which are obvious in calcium-dissolving ring, round in convex surface, fine, dense, white and aseptic mycelia, and observing the thalli by a microscope after gram staining for preliminary judgment.

(III) molecular biology identification of lactic acid bacteria for fermentation:

(l) Extracting a single-bacterium genome:

A. culturing the lactic acid bacteria obtained by screening in the step (II) overnight, taking l mL of the overnight-cultured bacterial suspension into a 1.5mL centrifuge tube, centrifuging at 10000rpm for 2min, and removing the supernatant to obtain thalli;

B. purging the thalli with l mL of sterile water, centrifuging at 10000rpm for 2min, and removing the supernatant to obtain the thalli;

C. adding 200 μ LSDS lysate, and water-bathing at 80 deg.C for 30 min;

D. adding 200 mu L of phenol-chloroform solution into the thallus lysate, wherein the composition and volume ratio of the phenol-chloroform solution are Tris saturated phenol: chloroform: isoamyl alcohol 25: 24: 1, reversing, uniformly mixing, centrifuging at 12000rpm for 5-10min, and taking 200 mu L of supernatant;

E. adding 400 μ L of glacial ethanol or glacial isopropanol into 200uL of supernatant, standing at-20 deg.C for 1h, centrifuging at 12000rpm for 5-10min, and discarding the supernatant;

F. adding 500 μ L70% (volume percentage) of glacial ethanol, resuspending the precipitate, centrifuging at 12000rpm for 1-3min, and discarding the supernatant;

drying in an oven at G.60 ℃ or naturally airing;

h.50. mu.L ddH2O pellet was re-solubilized for PCR;

(2)16S rDNA PCR

A. bacterial 16S rDNA 50 μ LPCR reaction:

10 × Taq buffer, 5 μ L; dNTP, 5. mu.L; 27F, 0.5 μ L; 1492R, 0.5 μ L; taq enzyme, 0.5. mu.L; template, 0.5 μ L; ddH2O, 38. mu.L.

PCR conditions:

95℃5min；95℃10s；55℃30s；72℃30s；step2-430×；72℃5min；12℃2min；

(3) preparing 1% agarose gel, mixing the PCR product with 10000 × loading buffer, loading the sample by 5 μ L, running at 120V for 30min, and performing gel imaging;

(4) and (3) carrying out sequencing analysis on the PCR product of the 16S rDNA, searching and similarity comparison on the obtained sequence result in GeneBank by using BLAST, and selecting a newly discovered strain with the sequencing result identified as belonging to the lactobacillus plantarum and preserving at-80 ℃ for later use.

Example 1: the lactobacillus plantarum CCFM1202 has good tolerance to simulated gastrointestinal fluid

Inoculating the lactobacillus plantarum CCFM1202 which is frozen and preserved in an MRS culture medium, carrying out anaerobic culture for 48h at the temperature of 37 ℃, carrying out subculture for 2-3 times by using the MRS culture medium, mixing 1mL of the culture medium of the lactobacillus plantarum CCFM1202 with 9.0mL of artificial simulated gastric juice (MRS culture medium containing 1% pepsin and pH 2.5), carrying out anaerobic culture at the temperature of 37 ℃, sampling at 0h, 0.5h, 1h and 2h respectively, carrying out pouring culture by using the MRS agar culture medium, carrying out plate colony counting, measuring the viable count and calculating the survival rate.

The survival rate is the ratio of the logarithmic viable count at the sampling time to the logarithmic viable count at the 0h time in the culture solution, and is expressed by%. Adding 1mL of culture solution of Lactobacillus plantarum CCFM1202 into 9mL of artificial simulated intestinal fluid (MRS culture medium containing 0.3% of bovine bile salt, 1% of trypsin and pH 8.0), anaerobically culturing at 37 ℃, sampling at 0h, 0.5h, 1h, 2h, 3h and 4h respectively, pouring and culturing by using MRS agar culture medium for counting plate colonies, measuring viable count and calculating survival rate. The survival rate is the ratio of the logarithmic viable count at the sampling time to the logarithmic viable count at the 0h time in the culture solution, and is expressed by%. The results of the experiment are shown in tables 1 and 2. The result shows that the lactobacillus plantarum CCFM1202 has better tolerance to the artificial gastrointestinal fluids.

TABLE 1 tolerance of Lactobacillus plantarum CCFM1202 in simulated gastric juice

TABLE 2 tolerance of Lactobacillus plantarum CCFM1202 in artificially simulated intestinal fluid

Example 2: lactobacillus plantarum CCFM1202 has no toxic and side effects on SD rats

Inoculating the freeze-preserved Lactobacillus plantarum CCFM1202 in MRS culture medium, anaerobically culturing at 37 deg.C for 48h, centrifuging to collect thallus, and suspending the Lactobacillus plantarum CCFM1202 thallus in sterile physiological saline to obtain a concentration of 3.0 × 10⁹CFU/mL of bacterial suspension.

8 healthy female SD rats with the weight of 200-.

The results of these tests are shown in Table 3. These results show that the feed concentration was 3.0X 10⁹The CFU/mL lactobacillus plantarum CCFM1202 has no obvious influence on rats, no obvious weight change and no death phenomenon. The appearance of the rat has no obvious pathological symptoms.

TABLE 3 weight change and death status of rats

Time (sky)	1	2	3	4	5	6	7
								Body weight (g)	203.1±2.5	205.2±3.1	206.6±3.7	207.7±4.2	208.8±4.6	209.7±3.5	211.3±5.3
Death situation	-	-	-	-	-	-	-

Note: -: no death of rat

Example 3: lactobacillus plantarum CCFM12022 has recovery effect on reproductive cycle disorder of polycystic ovarian syndrome rats

30 healthy female SD rats with the weight of 200-: the Lactobacillus plantarum of blank group (Control), Letrozole model group (Letrozole), Danine-35 group (Diane35), Lactobacillus plantarum CCFM1202 group (CCFM1202), Lactobacillus plantarum N34 group (N34, strain disclosed in Venus et al, having high cell adhesion and high biofilm formation ability, effectively inhibited the transcriptional activity of virulence factors of Campylobacter jejuni in mice [ J34 ]]Food and fermentation industry, 2020,46(14):12-18). Each group contains 6 rats with a gavage concentration of 3.0 × 10⁹CFU/mL of bacterial suspension 1 mL. The grouping and treatment methods of the experimental animals are shown in table 4:

TABLE 4 groups of experimental animals

Weeks 1-2: normal rats were gavaged with a 1% CMC solution and the remaining rats were gavaged with 1mg/kg/bw letrozole (suspended in 1% CMC) for modeling polycystic ovary syndrome. While processing was performed in the manner of table 4.

Weeks 3-4: the treatment was carried out in the manner of Table 4.

All experimental rats were examined daily for vaginal smears after the molding was stopped to observe changes in their reproductive cycle. The specific method comprises the following steps: the medical cotton swab is soaked in sterile normal saline, inserted into a rat vagina for about 1cm, gently rotated, and then taken out. Dropping a drop of sterile normal saline on a clean glass slide, lightly coating the sterile normal saline dropping area with a cotton swab taken out, and naturally drying. Staining the smear by using a Wright staining solution, observing the vaginal cell morphology under an optical microscope after the smear is dried, and recording the reproductive cycle, wherein the reproductive cycle is divided into an estrus interval, an early estrus stage, an estrus stage and a late estrus stage. Statistics were taken as the percentage of the interval between oestruses for the last five days for each rat.

Fresh excrement of rats is collected at the final stage of the test and is frozen at-80 ℃, the genome of the excrement is extracted, and the structure of intestinal flora is analyzed by using a second-generation sequencer. At the end of the test, rats are fasted for 12 hours without water prohibition, and are drunken by injecting a 1% sodium pentobarbital solution of 0.5mL/10g into the abdominal cavity, then the heart is sampled with blood, and the rats are killed by cervical dislocation. Centrifuging blood sample at 4 deg.C for 15min at 3000 Xg, collecting supernatant, and freezing at-80 deg.C for measuring related serum index. Collecting ovaries, rapidly placing in pre-cooled normal saline for rinsing to remove blood, placing in paraformaldehyde for fixation, and quickly freezing the remaining ovaries in liquid nitrogen and transferring to-80 ℃ for freezing and storing.

The estrus cycle is an important index for evaluating the sexual cycle of the rat, the estrus of a normal rat is 4-5 days, the prophase of the estrus, the anaphase of the estrus and the estrus interval repeatedly appear, and the disorder of the estrus cycle is an important characteristic of the PCOS rat. Table 5 is the effect of lactobacillus plantarum CCFM1202 on the reproductive cycle of polycystic ovary syndrome rats; table 5 records the change condition of the reproductive cycle of rats in different experimental groups on the last 5 days of animal experiments, the letrozole jelly mold enables the reproductive cycle of the rats to be always in the estrus period, and the ratio of the estrus period in the reproductive cycle is reduced by the intragastric lactobacillus plantarum CCFM1202, which shows that the lactobacillus plantarum CCFM1202 can restore the disorder of the reproductive cycle of the rats with polycystic ovary syndrome and has stronger effect than lactobacillus plantarum N34.

TABLE 5

Group of	Number of days in estrus interval (%)
		Control	30.00±10.95*
Letrozole	100.00±0.00
		Diane35	70.00±20.98*
CCFM1202	23.33±8.17*
		N34	96.67±8.17

Example 4: lactobacillus plantarum CCFM1202 can reduce testosterone level in rat serum with polycystic ovarian syndrome

The grouping, modeling and treatment methods of SD rats are the same as in example 3. At the end of the test, rats are fasted for 12 hours without water prohibition, and blood is collected from the heart after anesthesia by intraperitoneal injection of 0.5mL/10g of 1% sodium pentobarbital solution. Centrifuging blood sample at 3000 Xg and 4 deg.C for 10min, collecting supernatant, and determining testosterone content in serum according to the detection method of the kit.

The experimental result is shown in figure 2, the testosterone level in the serum of the rat of the letrozole molding group is obviously increased, and the intragastric lactobacillus plantarum CCFM1202 can obviously reduce the testosterone level in the serum of the rat. This shows that lactobacillus plantarum CCFM1202 can remarkably improve hyperandrogenism caused by polycystic ovary syndrome, and the effect is stronger than lactobacillus plantarum N34.

Example 5: lactobacillus plantarum CCFM1202 can reduce polycystic ovarian syndrome rat luteinizing hormone level

The SD rat grouping, modeling and treatment method are the same as example 3. At the end of the test, rats are fasted for 12 hours without water prohibition, and blood is collected from the heart after anesthesia by intraperitoneal injection of 0.5mL/10g of 1% sodium pentobarbital solution. Blood sample is centrifuged at 3000 Xg and 4 ℃ for 10min, the supernatant is taken, and the content of the luteinizing hormone in the serum is determined according to the detection method of the kit.

The experimental results are shown in figure 3. The luteinizing hormone level in the serum of the rat of the letrozole molding group is obviously increased, and the lactobacillus plantarum CCFM1202 in the gavage can obviously reduce the luteinizing hormone level in the serum of the rat. This shows that lactobacillus plantarum CCFM1202 can remarkably improve the level imbalance of luteinizing hormone caused by polycystic ovary syndrome, and the effect is stronger than lactobacillus plantarum N34.

Example 6: lactobacillus plantarum CCFM1202 can reduce the level of triglyceride in rat with polycystic ovarian syndrome

The SD rat grouping, modeling and treatment method are the same as example 3. At the end of the test, rats are fasted for 12 hours without water prohibition, and blood is collected from the heart after anesthesia by intraperitoneal injection of 0.5mL/10g of 1% sodium pentobarbital solution. Centrifuging blood sample at 3000 Xg and 4 deg.C for 10min, collecting supernatant, and determining triglyceride content in serum according to the detection method of the kit.

The experimental results are shown in figure 4. The level of triglyceride in the serum of a rat in a letrozole-made model is obviously increased, and the level of the triglyceride in the serum of the rat can be obviously reduced by the lactobacillus plantarum CCFM1202 with gavage. This shows that lactobacillus plantarum CCFM1202 can remarkably improve the level imbalance of triglyceride caused by polycystic ovary syndrome, and the effect is stronger than lactobacillus plantarum N34.

Example 7: lactobacillus plantarum CCFM1202 can reduce polycystic ovarian syndrome rat insulin resistance

The SD rat grouping, modeling and treatment method are the same as example 3. At the end of the test, rats are fasted for 12 hours without water prohibition, and blood is collected from the heart after anesthesia by intraperitoneal injection of 0.5mL/10g of 1% sodium pentobarbital solution. Centrifuging blood sample at 3000 Xg and 4 deg.C for 10min, collecting supernatant, and determining fasting blood glucose and fasting insulin level according to the detection method of the kit. The insulin resistance index is calculated from the fasting blood glucose and the fasting insulin level.

Insulin resistance index (fasting plasma glucose (mmol/L) × fasting insulin (mIU/L)/22.5

The experimental results are shown in figure 5. The insulin resistance index in the serum of the rat of the letrozole-made model group is obviously increased, and the insulin resistance index of the rat can be obviously reduced by the lactobacillus plantarum CCFM1202 with gavage. This shows that lactobacillus plantarum CCFM1202 can significantly improve insulin resistance caused by polycystic ovarian syndrome, and the action effect is stronger than lactobacillus plantarum N34.

Example 8: lactobacillus plantarum CCFM1202 can restore the reduction of intestinal flora diversity caused by polycystic ovarian syndrome

The grouping, modeling and treatment methods of SD rats are the same as in example 3. Towards the end of the experiment, fresh faeces were harvested from the rats and immediately frozen in a-80 ℃ freezer. And extracting a fecal genome, and analyzing the change of the intestinal flora by using a second-generation sequencing technology.

The flora analysis experiment result is shown in figure 6, the shannon index of a letrozole-made model rat is 5.80 +/-0.19, the shannon index of a CCFM1202 group is 6.14 +/-0.17, the shannon index of the intestinal flora of the rat with the polycystic ovary syndrome is obviously reduced, and the shannon index of the intestinal flora can be obviously adjusted back by taking the lactobacillus plantarum CCFM 1202. The result shows that the lactobacillus plantarum CCFM1202 screened by the invention has the capability of regulating the reduction of the intestinal flora diversity caused by the polycystic ovary syndrome, and the action effect is stronger than that of lactobacillus plantarum N34.

Example 9: lactobacillus plantarum CCFM1202 has strong in-vitro adsorption capacity on bisphenol F

The lactobacillus is purified and activated and cultured, inoculated in MRS liquid culture medium according to the inoculation amount of 1% (v/v), and cultured for 18h at 37 ℃. Then centrifuging at 8000r/min for 5min to collect thallus, collecting precipitate, cleaning with physiological saline, centrifuging at 8000r/min for 5min, and removing precipitate to obtain viable thallus cell, i.e. wet thallus. The wet cells were resuspended in 100mg/L of bisphenol F physiological saline to a final cell concentration of 1X 10⁹CFU/L. Culturing at 37 deg.C for 24h with 120r/min shaking table. Taking out the bisphenol F culture solution, centrifuging at 4 deg.C and 8000rpm for 10min, and transferring the supernatant for use.

And (4) carrying out HPLC sample injection pretreatment on the bisphenol F culture supernatant by adopting a solid-phase extraction mode. HPLC analysis conditions comprise that the model of the instrument is SHIMADZULC-20A, the detector is a two-stage array tube detector, the model of a chromatographic column is SinoChrom ODS-BP (4.6 multiplied by 250mm,5um), the mobile phase is methanol and water (7:3), the temperature of the column is 40 ℃, the flow rate is 0.5mL/min, the sample injection volume is 10uL, and the detection wavelength of BPF is 277 nm. And calculating the adsorption amount of the lactic acid bacteria to the bisphenol F according to the concentration difference of the bisphenol F before and after adsorption. The 3 replicates were averaged. The measurement result is shown in FIG. 7, the 24h in vitro adsorption rate of the Lactobacillus plantarum CCFM1202 on the bisphenol F reaches 60%, which is much higher than that of the control strain Lactobacillus plantarum N34.

Although the present invention has been described with reference to the preferred embodiments, it should be understood that various changes and modifications can be made therein by those skilled in the art without departing from the spirit and scope of the invention as defined in the appended claims.

Claims

1. A Lactobacillus plantarum (Lactobacillus plantarum) CCFM1202, which is deposited in Guangdong province microorganism strain collection center at 12 and 17 months in 2021, is deposited at Guangzhou microbial research institute of 5 th building, 59 th building, Middhou Zhou 100 th college, Middleya, Guangzhou city, and has a deposit number of GDMCC No: 62137.

2. a probiotic formulation comprising lactobacillus plantarum CCFM1202 as defined in claim 1.

3. The probiotic preparation according to claim 2, characterized in that the content of Lactobacillus plantarum CCFM1202 in the probiotic preparation is ≥ 1 x 10⁶CFU/mL or more than or equal to 1X 10⁶CFU/g。

4. A pharmaceutical composition comprising the lactobacillus plantarum CCFM1202 of claim 1.

5. The pharmaceutical composition of claim 4, further comprising a pharmaceutical carrier and/or a pharmaceutical excipient.

6. A fermented food, characterized in that it is produced by fermentation using the lactobacillus plantarum CCFM1202 according to claim 1.

7. The fermented food product according to claim 6, wherein the fermented food product comprises a solid food product, a liquid food product, a semi-solid food product.

8. Use of lactobacillus plantarum CCFM1202 as defined in claim 1 for the preparation of a product alleviating polycystic ovary syndrome.

9. Use according to claim 8, wherein the product has at least one of the following effects:

(1) the compound has better tolerance to simulated gastrointestinal fluid;

(3) significantly reducing the testosterone level of the rat with polycystic ovarian syndrome;

(5) significantly reducing the triglyceride level of a rat with polycystic ovarian syndrome;

(8) has obvious in-vitro adsorption capacity to bisphenol F.

10. The use according to claim 8, wherein the number of Lactobacillus plantarum CCFM1202 is ≥ 1 x 10 in said product⁶CFU/mL or more than or equal to 1X 10⁶CFU/g。